The AhpC and AhpD Antioxidant Defense System of Mycobacterium tuberculosis *

The peroxiredoxin AhpC from Mycobacterium tuberculosis has been expressed, purified, and characterized. It differs from other well characterized AhpC proteins in that it has three rather than one or two cysteine residues. Mutagenesis studies show that all three cysteine residues are important for catalytic activity. Analysis of the M. tuberculosis genome identified a second protein, AhpD, which has no sequence identity with AhpC but is under the control of the same promoter. This protein has also been cloned, expressed, purified, and characterized. AhpD, which has only been identified in the genomes of mycobacteria and Streptomyces viridosporus, is shown here to also be an alkylhydroperoxidase. The endogenous electron donor for catalytic turnover of the two proteins is not known, but both can be turned over with AhpF fromSalmonella typhimurium or, particularly in the case of AhpC, with dithiothreitol. AhpC and AhpD reduce alkylhydroperoxides more effectively than H2O2 but do not appear to interact with each other. These two proteins appear to be critical elements of the antioxidant defense system of M. tuberculosis and may be suitable targets for the development of novel anti-tuberculosis strategies.

[1]  T. Hakoshima,et al.  Crystal structure of a multifunctional 2-Cys peroxiredoxin heme-binding protein 23 kDa/proliferation-associated gene product. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Y. Nishiyama,et al.  Stimulation of peroxidase activity by decamerization related to ionic strength: AhpC protein from Amphibacillus xylanus. , 1999, Journal of biochemistry.

[3]  P. Ortiz de Montellano,et al.  Reduction of peroxides and dinitrobenzenes by Mycobacterium tuberculosis thioredoxin and thioredoxin reductase. , 1999, Archives of biochemistry and biophysics.

[4]  Alimuddin Zumla,et al.  Science, medicine, and the future: Tuberculosis , 1998 .

[5]  B. Barrell,et al.  Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence , 1998, Nature.

[6]  David A. Mead,et al.  Inhibition of a Mycobacterium tuberculosis β-Ketoacyl ACP synthase by isoniazid , 1998 .

[7]  J. Evans.  TB : KNOW YOUR ENEMY , 1998 .

[8]  R. Slayden,et al.  Mechanisms of isoniazid resistance in Mycobacterium tuberculosis. , 1998, Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy.

[9]  L. Poole,et al.  Novel application of 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole to identify cysteine sulfenic acid in the AhpC component of alkyl hydroperoxide reductase. , 1997, Biochemistry.

[10]  L. Poole,et al.  Requirement for the two AhpF cystine disulfide centers in catalysis of peroxide reduction by alkyl hydroperoxide reductase. , 1997, Biochemistry.

[11]  L. Poole,et al.  Roles for the two cysteine residues of AhpC in catalysis of peroxide reduction by alkyl hydroperoxide reductase from Salmonella typhimurium. , 1997, Biochemistry.

[12]  S. Dhandayuthapani,et al.  Molecular basis for the exquisite sensitivity of Mycobacterium tuberculosis to isoniazid. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[13]  S. Dhandayuthapani,et al.  The extreme sensitivity of Mycobacterium tuberculosis to the front-line antituberculosis drug isoniazid , 1996, Nature Biotechnology.

[14]  D. Sherman,et al.  Biochemical and genetic data suggest that InhA is not the primary target for activated isoniazid in Mycobacterium tuberculosis. , 1996, The Journal of infectious diseases.

[15]  Joanne I. Yeh,et al.  Structure of the native cysteine-sulfenic acid redox center of enterococcal NADH peroxidase refined at 2.8 A resolution. , 1996, Biochemistry.

[16]  Clifton E. Barry,et al.  Compensatory ahpC Gene Expression in Isoniazid-Resistant Mycobacterium tuberculosis , 1996, Science.

[17]  W. Adam,et al.  The Schenck Ene Reaction: Diastereoselective Oxyfunctionalization with Singlet Oxygen in Synthetic Applications†‡ , 1996 .

[18]  M. Fenton,et al.  Immunopathology of tuberculosis: roles of macrophages and monocytes , 1996, Infection and immunity.

[19]  L. Poole.  Flavin-dependent alkyl hydroperoxide reductase from Salmonella typhimurium. 2. Cystine disulfides involved in catalysis of peroxide reduction. , 1996, Biochemistry.

[20]  L. Poole,et al.  Flavin-dependent alkyl hydroperoxide reductase from Salmonella typhimurium. 1. Purification and enzymatic activities of overexpressed AhpF and AhpC proteins. , 1996, Biochemistry.

[21]  T. Ottenhoff,et al.  Identification and functional characterization of thioredoxin of Mycobacterium tuberculosis , 1995, Infection and immunity.

[22]  C. Pace,et al.  How to measure and predict the molar absorption coefficient of a protein , 1995, Protein science : a publication of the Protein Society.

[23]  R. Offringa,et al.  Purification and Functional Analysis of the Mycobacterium leprae Thioredoxin/Thioredoxin Reductase Hybrid Protein (*) , 1995, The Journal of Biological Chemistry.

[24]  G. Mahairas,et al.  Disparate responses to oxidative stress in saprophytic and pathogenic mycobacteria. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[25]  S. Cole,et al.  Missense mutations in the catalase‐peroxidase gene, katG, are associated with isoniazid resistance in Mycobacterium tuberculosis , 1995, Molecular microbiology.

[26]  S. Rhee,et al.  Thioredoxin-dependent peroxide reductase from yeast. , 1994, The Journal of biological chemistry.

[27]  P. Schultz,et al.  Mechanistic Studies of the Oxidation of Isoniazid by the Catalase Peroxidase from Mycobacterium tuberculosis , 1994 .

[28]  G. Church,et al.  Cloning and sequencing of thiol-specific antioxidant from mammalian brain: alkyl hydroperoxide reductase and thiol-specific antioxidant define a large family of antioxidant enzymes. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[29]  W. Jacobs,et al.  inhA, a gene encoding a target for isoniazid and ethionamide in Mycobacterium tuberculosis. , 1994, Science.

[30]  S. Kaufmann,et al.  Immune Mechanisms of Protection , 1994 .

[31]  S T Cole,et al.  Characterization of the katG gene encoding a catalase-peroxidase required for the isoniazid susceptibility of Mycobacterium tuberculosis , 1993, Journal of bacteriology.

[32]  D. Young,et al.  Transformation with katG restores isoniazid‐sensitivity in Mycobacterium tuberculosis isolates resistant to a range of drug concentrations , 1993, Molecular microbiology.

[33]  W. Roper,et al.  The new tuberculosis. , 1992, The New England journal of medicine.

[34]  A. G. Davies,et al.  The mechanisms of the rearrangements of allylic hydroperoxides: 5α-hydroperoxy-3β-hydroxycholest-6-ene and 7α-hydroperoxy-3β-hydroxycholest-5-ene , 1989 .

[35]  N. Porter,et al.  Allylic hydroperoxide rearrangement. .beta.-Scission or concerted pathway? , 1987 .

[36]  H. Wasserman,et al.  Singlet oxygen in organic synthesis , 1981 .

[37]  M. Maumy,et al.  Intervention d'un mecanisme ionique dans la photooxygenation sensibilisee de l′α-pinene en milieu protique , 1980 .

[38]  K. S. Iyer,et al.  Direct spectrophotometric measurement of the rate of reduction of disulfide bonds. The reactivity of the disulfide bonds of bovine -lactalbumin. , 1973, The Journal of biological chemistry.

[39]  A. Nickon,et al.  Reactivity and Geometry in Allylic Systems. I. Stereochemistry of Photosensitized Oxygenation of Monoölefins1,2 , 1961 .

[40]  G. Schenck,et al.  Photochemische Reaktionen III. Über die Bildung von Hydroperoxyden bei photosensibilisierten Reaktionen von O2 mit geeigneten Akzeptoren, insbesondere mit α- und β-Pinen , 1953 .